共培养下后交叉韧带成纤维细胞中赖氨酰氧化酶的基因表达**☆

摘要

背景：后交叉韧带对人膝关节结构的稳定以及功能的发挥起着重要作用。与内侧副韧带相比，损伤后的后交叉韧带难以很好的自我愈合，甚至会导致半月板撕裂和软骨损伤。为了提高后交叉韧带的愈合能力，这就需要寻找新的途径来再生和修复损伤的后交叉韧带。近年来的研究表明，赖氨酰氧化酶在组织损伤修复过程中起到非常重要的作用，但其在后交叉韧带修复过程中的分子机制研究尚未涉猎。目的：观察与滑膜细胞共培养后交叉韧带成纤维细胞中赖氨酰氧化酶基因的表达。方法：将第4代的后交叉韧带成纤维细胞和滑膜细胞分别种植于6孔板和Tanswel 中。实验分为2组，即后交叉韧带成纤维细胞与滑膜细胞共培养组和后交叉韧带成纤维细胞单层培养组。培养6 h后，提取总RNA，通过半定量PCR和实时定量PCR检测单层培养组和共培养组中后交叉韧带成纤维细胞中赖氨酰氧化酶基因表达。结果与结论：与单层培养组相比，赖氨酰氧化酶、赖氨酰氧化酶样蛋白1、赖氨酰氧化酶样蛋白2、赖氨酰氧化酶样蛋白3和赖氨酰氧化酶样蛋白4的基因表达在共培养的后交叉韧带细胞中都明显升高，分别增加了1.1，1.4，1.1，1.3，1.1倍(P <0.05)。单层培养组家族成员在单层培养和共培养中表达情况的差异性，说明细胞之间的相互作用会影响后交叉韧带组织的损伤修复，对后交叉韧带损伤修复的机制研究有极其重要的参考价值。%BACKGROUND: The posterior cruciate ligament is an important ligament contributing to the stability and normal functioning of the knee joint. Compared to the medial col ateral ligament, the posterior cruciate ligament has a poor healing ability and often leads to significant knee instability and secondary knee damage including meniscus tears and articular cartilage injuries. In attempting to improve the healing ability of injured posterior cruciate ligament, we need to find new ways for regeneration and repair of injured posterior cruciate ligament. Previous studies have demonstrated that lysyl oxidases play an important role in the tissue repair mechanism, but the effect of lysyl oxidases from injured posterior cruciate ligament on the process of wound repair remains unclear. OBJECTIVE: To investigate the expressions of lysyl oxidases in the posterior cruciate ligament fibroblasts co-cultured with synovial cel s. METHODS: The fourth passage of posterior cruciate ligament fibroblasts and synovial cel s were placed in 6-wel plates and Transwel , respectively. Two groups were designed as fol ows, posterior cruciate ligament fibroblasts group as control and co-cultured group termed as test group. At 6 hours after co-culture, total RNA was isolated and the expressions of lysyl oxidases in the posterior cruciate ligament fibroblasts were analyzed by semi-quantitative reverse-transcription PCR and quantitative real-time PCR. RESULTS AND CONCLUSION: The results revealed that co-culture contributed to up-regulations of lysyl oxidases compared with the control group, and gene levels were up to 1.1 folds in lysyl oxidase, 1.4 folds in lysyl oxidase-like 1 protein, 1.1 folds in lysyl oxidase-like 2 protein, 1.3 folds in lysyl oxidase-like 3 protein, 1.1 folds in lysyl oxidase-like 4 protein in co-cultured posterior cruciate ligament cel s (P < 0.05). The differential expression of lysyl oxidases in co-cultured posterior cruciate ligament cel s implies that cel-cel interaction and crosstalk are related with posterior cruciate ligament wound healing and have significant potential value and clinical usage for cure of injured posterior cruciate ligament.